Analyzing the factors affecting virus invasion by quantitative single-particle analysis.

Viral diseases are among the main threats to public health. Understanding the factors affecting viral invasion is important for antiviral research. Until now, it was known that most viruses have very low plaque-forming unit (PFU)-to-particle ratios. However, further investigation is required to determine the underlying factors. Here, using quantitative single-particle analysis methods, the invasion of Semliki Forest virus (SFV), Japanese encephalitis virus (JEV), and influenza A virus (IAV) containing attachment to the cell surface, entry into the cell, transport towards the cell interior, and fusion with endosomes to release nucleocapsids were quantitatively analysed in parallel. It was found that for SFV with an PFU-to-particle ratio of approximately 1:2, an entry efficiency of approximately 31% limited infection. For JEV, whose PFU-to-particle ratio was approximately 1:310, an attachment efficiency of approximately 27% and an entry efficiency of 10% were the main factors limiting its infection. Meanwhile, for IAV with PFU-to-particle ratios of 1:8100, 5% attachment efficiency, 9% entry efficiency, and 53% fusion efficiency significantly limited its infection. These results suggest that viruses with different infectivities have different limited steps in the invasion process. Moreover, there are significant differences in attachment efficiencies among viruses, emphasizing the pivotal role of attachment in viral invasion. The influence of the virus purification method on virus invasion was also investigated. This study, for the first time, reports the efficiencies of different stages of virus invasion, leading to a better understanding of virus invasion and providing a protocol to quantitatively analyse the virus invasion efficiency.

Localized Path Planning for Mobile Robots Based on a Subarea-Artificial Potential Field Model.

The artificial potential field method has efficient obstacle avoidance ability, but this traditional method suffers from local minima, unreasonable paths, and sudden changes in heading angles during obstacle avoidance, leading to rough paths and increased energy consumption. To enable autonomous mobile robots (AMR) to escape from local minimum traps and move along reasonable, smooth paths while reducing travel time and energy consumption, in this paper, an artificial potential field method based on subareas is proposed. First, the optimal virtual subgoal was obtained around the obstacles based on the relationship between the AMR, obstacles, and goal points in the local environment. This was done according to the virtual subgoal benefit function to solve the local minima problem and select a reasonable path. Secondly, when AMR encountered an obstacle, the subarea-potential field model was utilized to solve problems such as path zigzagging and increased energy consumption due to excessive changes in the turning angle; this helped to smooth its planning path. Through simulations and actual testing, the algorithm in this paper demonstrated smoother heading angle changes, reduced energy consumption, and a 10.95% average reduction in movement time when facing a complex environment. This proves the feasibility of the algorithm.

Contralateral renal change in a unilateral ureteral obstruction rat model using intravoxel incoherent motion diffusion-weighted imaging.

OBJECTIVES: Most functional magnetic resonance research has primarily examined alterations in the affected kidney, often neglecting the contralateral kidney. Our study aims to investigate whether imaging parameters accurately depict changes in both the renal cortex and medulla in a unilateral ureteral obstruction rat model, thereby showcasing the utility of intravoxel incoherent motion (IVIM) in evaluating contralateral renal changes. METHODS: Six rats underwent MR scans and were subsequently sacrificed for baseline histological examination. Following the induction of left ureteral obstruction, 48 rats were scanned, and the histopathological examinations were conducted on days 3, 7, 10, 14, 21, 28, 35, and 42. The apparent diffusion coefficient (ADC), pure molecular diffusion (D), pseudodiffusion (D*), and perfusion fraction (f) values were measured using IVIM. RESULTS: On the 10th day of obstruction, both cortical and medullary ADC values differed significantly between the UUO10 group and the sham group (p < 0.01). The cortical D values showed statistically significant differences between UUO3 group and sham group (p < 0.01) but not among UUO groups at other time point. Additionally, the cortical and medullary f values were statistically significant between the UUO21 group and the sham group (p < 0.01). Especially, the cortical f values exhibited significant differences between the UUO21 group and the UUO groups with shorter obstruction time (at time point of 3, 7, 10, 14 day) (p < 0.01). CONCLUSIONS: Significant hemodynamic alterations were observed in the contralateral kidney following renal obstruction. IVIM accurately captures changes in the unobstructed kidney. Particularly, the cortical f value exhibits the highest potential for assessing contralateral renal modifications.

All-in-one bio-derived poly(L-lactic acid)-based composite with fire-resistance and smoke-suppression performance.

The flammability of bio-derived poly(L-lactic acid) (PLA) greatly limits its application and eco-friendly multifunctional fire-fighting PLA-based composites are highly desired. In this work, a fully bio-based modified CS (C-CS) and commercially available eco-friendly ammonium polyphosphate (APP) were used as a synergistic flame retardant agent (C-CS/APP) to investigate its effects on fire-proofing performance and diverse properties of the PLA. The PLA/5%C-CS/5%APP composite exhibited excellent fire-resistant performance with anti-droplet, smoke-suppression and self-extinguishing property, and its limited oxygen index enhanced by 37 % (compared with neat PLA). This composite reached the highest V-0 fire safety rating, and its peak of heat release rate and total smoke production reduced by 26.5 % and 68.3 %, respectively. In addition, the char residue yield after the cone calorimeter test increased by 46 times in the composite, indicating an outstanding char-forming capacity. The condensed phase flame retardancy played a crucial role on the fire-fighting of this composite, that is, significantly enhanced char residue (as a physical barrier) blocked the heat exchange and O2 entry, and further suppressed the combustion reaction. Additionally, the PLA-based composite showed outstanding UV-absorption property, good anti-bacterial effect, and increased hydrophilicity and crystallizability.

Nondestructive Determination of Epicarp Hardness of Passion Fruit Using Near-Infrared Spectroscopy during Storage.

The hardness of passion fruit is a critical feature to consider when determining maturity during post-harvest storage. The capacity of near-infrared diffuse reflectance spectroscopy (NIRS) for non-destructive detection of outer and inner hardness of passion fruit epicarp was investigated in this work. The passion fruits' spectra were obtained using a near-infrared spectrometer with a wavelength range of 10,000-4000 cm-1. The hardness of passion fruit's outer epicarp (F1) and inner epicarp (F2) was then measured using a texture analyzer. Moving average (MA) and mean-centering (MC) techniques were used to preprocess the collected spectral data. Competitive adaptive reweighted sampling (CARS), successive projection algorithm (SPA), and uninformative variable elimination (UVE) were used to pick feature wavelengths. Grid-search-optimized random forest (Grids-RF) models and genetic-algorithm-optimized support vector regression (GA-SVR) models were created as part of the modeling process. After MC preprocessing and CARS selection, MC-CARS-Grids-RF model with 7 feature wavelengths had the greatest prediction ability for F1. The mean square error of prediction set (RMSEP) was 0.166 gN. Similarly, following MA preprocessing, the MA-Grids-RF model displayed the greatest predictive performance for F2, with an RMSEP of 0.101 gN. When compared to models produced using the original spectra, the R2P for models formed after preprocessing and wavelength selection improved. The findings showed that near-infrared spectroscopy may predict the hardness of passion fruit epicarp, which can be used to identify quality during post-harvest storage.

Prokaryotic expression, purification, and the in vitro and in vivo protection study of dehydrin WDHN2 from Triticum aestivum.

Dehydrins proteins accumulate and play important protective roles in most plants during abiotic stresses. The objective of this study was to characterize a YSK2-type dehydrin gene, WDHN2, isolated from Triticum aestivum previously. In this work, wheat dehydrin WDHN2 was expressed in Escherichia coli and purified by immobilized metal affinity chromatography, which exhibited as a single band by sodium dodecyl sulfonate polyacrylamide gel electrophoresis and western blotting. We show that WDHN2 is capable of alleviating lactate dehydrogenase inactivation from heat and desiccation in vitro enzyme activity protection assay. In vivo assay of Escherichia coli viability demonstrates the enhancement of cell survival by the overexpression of WDHN2. The protein aggregation prevention assay explores that WDHN2 has a broad protective effect on the cellular proteome. The results show that WDHN2 is mainly accumulated in the nucleus and cytosol, suggesting that this dehydrin may exert its function in both cellular compartments. Our data suggest that WDHN2 acts as a chaperone molecular in vivo.

One-Step Dual-Color Labeling of Viral Envelope and Intraviral Genome with Quantum Dots Harnessing Virus Infection.

Labeling the genome and envelope of a virus with multicolor quantum dots (QDs) simultaneously enables real-time monitoring of viral uncoating and genome release, contributing to our understanding of virus infection mechanisms. However, current labeling techniques require genetic modification, which alters the virus's composition and infectivity. To address this, we utilized the CRISPR/Cas13 system and a bioorthogonal metabolic method to label the Japanese encephalitis virus (JEV) genome and envelopes with different-colored QDs in situ. This technique allows one-step two-color labeling of the viral envelope and intraviral genome with QDs harnessing virus infection. In combination with single-virus tracking, we visualized JEV uncoating and genome release in real time near the endoplasmic reticulum of live cells. This labeling strategy allows for real-time visualization of uncoating and genome release at the single-virus level, and it is expected to advance the study of other viral infection mechanisms.

Outer membrane vesicles of avian pathogenic Escherichia coli induce necroptosis and NF-κB activation in chicken macrophages via RIPK1 mediation.

Outer membrane vesicles (OMVs) are soluble mediators secreted by Gram-negative bacteria that are involved in communication. They can carry a variety of harmful molecules, which induce cytotoxic responses and inflammatory reactions in the absence of direct host cell-bacterium interactions. We previously reported the isolation of OMVs from avian pathogenic Escherichia coli (APEC) culture medium by ultracentrifugation, and characterized them as a substance capable of inducing the production of pro-inflammatory cytokines and causing tissue damage. However, the specific mechanisms by which APEC-secreted OMVs activate host cell death signaling and inflammation are poorly understood. Here, we show that OMVs are involved in the pathogenesis of APEC disease. In an APEC/chicken macrophage (HD11) coculture system, APEC significantly promoted HD11 cell death and inflammatory responses by secreting OMVs. Using western blotting analysis and specific pathway inhibitors, we demonstrated that the induction of HD11 death by APEC OMVs is associated with the activation of receptor interacting serine/threonine kinase 1 (RIPK1)-, receptor interacting serine/threonine kinase 3 (RIPK3)-, and mixed lineage kinase like pseudokinase (MLKL)-induced necroptosis. Notably, necroptosis inhibitor-1 (Nec-1), an RIPK1 inhibitor, reversed these effects. We also showed that APEC OMVs promote the activation of the NF-κB signaling pathway, leading to the phosphorylation of IκB-α and p65, the increased nuclear translocation of p65, and the significant upregulation of interleukin 1ß (IL-1ß) and IL-6 transcription. Importantly, APEC OMVs-induced IL-1ß and IL-6 mRNA expression and the activation of the NF-κB signaling pathway were similarly significantly inhibited by a RIPK1-specific inhibitor. Based on these findings, we have established that RIPK1 plays a dual role in HD11 cells necroptosis and the proinflammatory cytokine (IL-1ß and IL-6) expression induced by APEC OMVs. RIPK1 mediated the induction of necroptosis and the activation of the NF-κB in HD11 cells via APEC OMVs. The results of this study provide a basis for further investigation of the contribution of OMVs to the pathogenesis of APEC.

Real-Time Dissection of the Exosome Pathway for Influenza Virus Infection.

Exosomes play an important role in the spread of viral infections and immune escape. However, the exact ability and mechanisms by which exosomes produced during viral infections (vExos) infect host cells are still not fully understood. In this study, we developed a dual-color exosome labeling strategy that simultaneously labels the external and internal structures of exosomes with quantum dots to enable in situ monitoring of the transport process of vExos in live cells using the single-particle tracking technique. Our finding revealed that vExos contains the complete influenza A virus (IAV) genome and viral ribonucleoprotein complexes (vRNPs) proteins but lacks viral envelope proteins. Notably, these vExos have the ability to infect cells and produce progeny viruses. We also found that vExos are transported in three stages, slow-fast-slow, and move to the perinuclear region via microfilaments and microtubules. About 30% of internalized vExos shed the external membrane and release the internal vRNPs into the cytoplasm by fusion with endolysosomes. This study suggested that vExos plays a supporting role in IAV infection by assisting with IAV propagation in a virus-independent manner. It emphasizes the need to consider the infectious potential of vExos and draws attention to the potential risk of exosomes produced by viral infections.

Mucoepidermoid carcinoma of the lung with hemoptysis as initial symptom: A case report.

BACKGROUND: Mucoepidermoid carcinoma of the lung is a rare malignant tumor, accounting for 0.1%-0.2% of all lung malignancies. It is a primary salivary gland tumor of the lung. Surgical resection is the primary treatment for pulmonary mucoepidermoid carcinoma, for which there has been no standardized treatment strategy. This article reports a case of a young woman with pulmonary mucoepidermoid carcinoma with hemoptysis as the first symptom. CASE SUMMARY: A 24-year-old female patient presented with "4 d of hemoptysis" as the chief complaint. She had no special history and denied any smoking or drinking history. Physical examination revealed that the vital signs were stable and scattered small wet rales were heard in the left lung. After admission, the lung tumor markers were checked, and no abnormalities were found. After completing the bronchoscopy, a spherical lesion was observed at the main bronchus 1.5 cm away from the protubercle, with obvious pulsation and little blood seepage on the surface, and histopathological biopsy results showed acute and chronic inflammation. She was transferred to the Department of Thoracic Surgery for surgical treatment on the 16th day after admission. After exclusion of surgical conjunctures, the patient underwent resection of the tumor in the left main bronchus with single-pore video-assisted thoracic surgery on the 19th day after admission. The postoperative histopathological biopsy results showed mucoepidermoid carcinoma of the lung. The patient and her family refused to complete genetic testing and she was discharged from the hospital on the 8th day after surgery. During the follow-up period, the patient experienced shortness of breath after feeling active and had no special discomfort. CONCLUSION: We have documented a case of moderately differentiated mucoepidermoid lung cancer with hemoptysis as the first symptom to improve clinicians' understanding of the disease and provide a new dimension of thinking for its future diagnosis and treatment.

Staphylococcus warneri strain XSB102 exacerbates psoriasis and promotes keratinocyte proliferation in imiquimod-induced psoriasis-like dermatitis mice.

Psoriasis is one of the common chronic inflammatory skin diseases worldwide. The skin microbiota plays a role in psoriasis through regulating skin homeostasis. However, the studies on the interactions between symbiotic microbial strains and psoriasis are limited. In this study, Staphylococcus strain XSB102 was isolated from the skin of human, which was identified as Staphylococcus warneri using VITEK2 Compact. To reveal the roles of Staphylococcus warneri on psoriasis, XSB102 were applied on the back of imiquimod-induced psoriasis-like dermatitis mice. The results indicated that it exacerbated the psoriasis and significantly increased the thickening of the epidermis. Furthermore, in vitro experiments confirmed that inactivated strain XSB102 could promote the proliferation of human epidermal keratinocytes (HaCaT) cell. However, real-time quantitative PCR and immunofluorescence results suggested that the expression of inflammatory factors such as IL-17a, IL-6, and so on were not significantly increased, while extracellular matrix related factors such as Col6a3 and TGIF2 were significantly increased after XSB102 administration. This study indicates that Staphylococcus warneri XSB102 can exacerbate psoriasis and promote keratinocyte proliferation independently of inflammatory factors, which paves the way for further exploration of the relationship between skin microbiota and psoriasis.

Research on automatic classification and detection of chicken parts based on deep learning algorithm.

Accurate classification and identification of chicken parts are critical to improve the productivity and processing speed in poultry processing plants. However, the overlapping of chicken parts has an impact on the effectiveness of the identification process. To solve this issue, this study proposed a real-time classification and detection method for chicken parts, utilizing YOLOV4 deep learning. The method can identify segmented chicken parts on the assembly line in real time and accurately, thus improving the efficiency of poultry processing. First, 600 images containing multiple chicken part samples were collected to build a chicken part dataset after using the image broadening technique, and then the dataset was divided according to the 6:2:2 division principle, with 1200 images as the training set, 400 images as the test set, and 400 images as the validation set. Second, we utilized the single-stage target detector YOLO to predict and calculate the chicken part images, obtaining the categories and positions of the chicken leg, chicken wing, and chicken breast in the image. This allowed us to achieve real-time classification and detection of chicken parts. This approach enabled real-time and efficient classification and detection of chicken parts. Finally, the mean average precision (mAP) and the processing time per image were utilized as key metrics to evaluate the effectiveness of the model. In addition, four other target detection algorithms were introduced for comparison with YOLOV4-CSPDarknet53 in this study, which include YOLOV3-Darknet53, YOLOV3-MobileNetv3, SSD-MobileNetv3, and SSD-VGG16. A comprehensive comparison test was conducted to assess the classification and detection performance of these models for chicken parts. Finally, for the chicken part dataset, the mAP of the YOLOV4-CSPDarknet53 model was 98.86% on a single image with an inference speed of 22.2 ms, which was higher than the other four models of YOLOV3-Darknet53, YOLOV3-MobileNetv3, SSD-MobileNetv3, and SSD-VGG16 mAP by 3.27%, 3.78%, 6.91%, and 6.13%, respectively. The average detection time was reduced by 13, 1.9, 6.2, and 20.3 ms, respectively. In summary, the chicken part classification and detection method proposed in this study offers numerous benefits, including the ability to detect multiple chicken parts simultaneously, as well as delivering high levels of accuracy and speed. Furthermore, this approach effectively addresses the issue of accurately identifying individual chicken parts in the presence of occlusion, thereby reducing waste on the assembly line. PRACTICAL APPLICATION: The aim of this study is to offer visual technical assistance in minimizing wastage and resource depletion during the sorting and cutting of chicken parts in poultry production and processing facilities. Furthermore, considering the diverse demands and preferences regarding chicken parts, this research can facilitate product processing that caters specifically to consumer preferences.

EspE3 plays a role in the pathogenicity of avian pathogenic Escherichia coli.

APEC encodes multiple virulence factors that have complex pathogenic mechanisms. In this study, we report a virulence factor named EspE3, which can be secreted from APEC. This protein was predicted to have a leucine-rich repeat domain (LRR) and may have a similar function to IpaH class effectors of the type III secretion system (T3SS). For further exploration, the regulatory correlation between the espE3 and ETT2 genes in APEC was analysed. We then assessed the pathogenicity of EspE3, detected it in APEC secretion proteins and screened the proteins of EspE3 that interact with chicken trachea epithelial cells. This study provides data on a new virulence factor for further exploring the pathogenic mechanism of APEC.

Anti-interleukin 33 treatment alleviates psoriatic dermatitis in mice induced imiquimod.

Interleukin-33（IL-33), is constitutively expressed in the epithelial cells of the skin. It has been reported that IL-33 contributed to the severity of the disease in psoriasis-like mouse models. In the current study, we evaluated the effect of anti-IL-33 antibody (Ab) in imiquimod-induced psoriatic dermatitis in mice. Our observations showed that anti-IL-33 Ab ameliorated the erythema, scaling, epidermal thickness and spleen index. Additionally, we found anti-IL-33 Ab significantly decreased the expression of psoriasis-related cytokines. Moreover, anti-IL-33 Ab significantly reduced Ki-67 positive cells, CD3+CD4+T cells, and CD3+CD8+T cells in the skin lesions. Furthermore, anti-IL-33 Ab treatment down-regulated the expression of phosphorylation of STAT3 and IL-33 in model mouse. These results indicated that the anti-IL-33 Ab alleviated the seriousness of skin lesions, inhibited the activation of the STAT3, lymphocyte infiltration and the secretion of pro-inflammatory cytokines in imiquimod-induced psoriatic dermatitis in mice, suggesting IL-33 may be a therapeutic target for the treatment of psoriasis.

Research on Online Nondestructive Detection Technology of Duck Egg Origin Based on Visible/Near-Infrared Spectroscopy.

As living standards rise, people have higher requirements for the quality of duck eggs. The quality of duck eggs is related to their origin. Thus, the origin traceability and identification of duck eggs are crucial for protecting the rights and interests of consumers and preserving food safety. As the world's largest producer and consumer of duck eggs, China's duck egg market suffers from a severe lack of duck egg traceability and rapid origin identification technology. As a result, a large number of duck eggs from other regions are sold as products from well-known brands, which seriously undermines the rights and interests of consumers and is not conducive to the sound development of the duck egg industry. To address the above issues, this study collected visible/near-infrared spectral data online from duck eggs of three distinct origins. To reduce noise in the spectral data, various pre-processing algorithms, including MSC, SNV, and SG, were employed to process the spectral data of duck eggs in the range of 400-1100 nm. Meanwhile, CARS and SPA were used to select feature variables that reflect the origin of duck eggs. Finally, classification models of duck egg origin were developed based on RF, SVM, and CNN, achieving the highest accuracy of 97.47%, 98.73%, and 100.00%, respectively. To promote the technology's implementation in the duck egg industry, an online sorting device was built for duck eggs, which mainly consists of a mechanical drive device, spectral software, and a control system. The online detection performance of the machine was validated using 90 duck eggs, and the final detection accuracy of the RF, SVM, and CNN models was 90%, 91.11%, and 94.44%, with a detection speed of 0.1 s, 0.3 s, and 0.5 s, respectively. These results indicate that visible/near-infrared spectroscopy can be exploited to realize rapid online detection of the origin of duck eggs, and the methodologies used in this study can be immediately implemented in production practice.

The transcriptional regulator EtrA mediates ompW contributing to the pathogenicity of avian pathogenic Escherichia coli.

Avian pathogenic Escherichia coli (APEC) causes avian colibacillosis and leads to high mortality in poultry and huge economic losses. Therefore, it is important to investigate the pathogenic mechanisms of APEC. Outer membrane protein OmpW is involved in the environmental adaptation and pathogenesis of Gram-negative bacteria. OmpW is regulated by many proteins, including FNR, ArcA, and NarL. In previous studies, regulator EtrA is involved in the pathogenicity of APEC and affects the transcript levels of ompW. However, the function of OmpW in APEC and its regulation remain unclear. In this study, we constructed mutant strains with altered etrA and/or ompW genes to evaluate the roles of EtrA and OmpW in the biological characteristics and pathogenicity of APEC. Compared with wild-type strain AE40, mutant strains ∆etrA, ∆ompW, and ∆etrA∆ompW showed significantly lower motility, lower survival under external environmental stress, and lower resistance to serum. Biofilm formation by ∆etrA and ∆etrA∆ompW was significantly enhanced relative to that of AE40. The transcript levels of TNF-α, IL1ß, and IL6 were also significantly enhanced in DF-1 cells infected with these mutant strains. Animal infection assays showed that deletion of etrA and ompW genes attenuated the virulence of APEC in chick models, and damage to the trachea, heart, and liver caused by these mutant strains was attenuated relative to that caused by the wild-type strain. RT-qPCR and ß-galactosidase assay showed that EtrA positively regulates the expression of the ompW gene. These findings demonstrate that regulator EtrA positively regulates the expression of OmpW, and that they both contribute to APEC motility, biofilm formation, serum resistance, and pathogenicity.

Real-Time Dissection of the Transportation and miRNA-Release Dynamics of Small Extracellular Vesicles.

Extracellular vesicles (EVs) are cell-derived membrane-enclosed structures that deliver biomolecules for intercellular communication. Developing visualization methods to elucidate the spatiotemporal dynamics of EVs' behaviors will facilitate their understanding and translation. With a quantum dot (QD) labeling strategy, a single particle tracking (SPT) platform is proposed here for dissecting the dynamic behaviors of EVs. The interplays between tumor cell-derived small EVs (T-sEVs) and endothelial cells (ECs) are specifically investigated based on this platform. It is revealed that, following a clathrin-mediated endocytosis by ECs, T-sEVs are transported to the perinuclear region in a typical three-stage pattern. Importantly, T-sEVs frequently interact with and finally enter lysosomes, followed by quick release of their carried miRNAs. This study, for the first time, reports the entire process and detailed dynamics of T-sEV transportation and cargo-release in ECs, leading to better understanding of their proangiogenic functions. Additionally, the QD-based SPT technique will help uncover more secrets of sEV-mediated cell-cell communication.

The two-component system histidine kinase EnvZ contributes to Avian pathogenic Escherichia coli pathogenicity by regulating biofilm formation and stress responses.

EnvZ, the histidine kinase (HK) of OmpR/EnvZ, transduces osmotic signals in Escherichia coli K12 and affects the pathogenicity of Shigella flexneri and Vibrio cholera. Avian pathogenic E. coli (APEC) is an extra-intestinal pathogenic E. coli (ExPEC), causing acute and sudden death in poultry and leading to severe economic losses to the global poultry industry. How the functions of EnvZ correlate with APEC pathogenicity was still unknown. In this study, we successfully constructed the envZ mutant strain AE17ΔenvZ and the inactivation of envZ significantly reduced biofilms and altered red, dry, and rough (rdar) morphology. In addition, AE17ΔenvZ was significantly less resistant to acid, alkali, osmotic, and oxidative stress conditions. Deletion of envZ significantly enhanced sensitivity to specific pathogen-free (SPF) chicken serum and increased adhesion to chicken embryonic fibroblast DF-1 cells and elevated inflammatory cytokine IL-1ß, IL6, and IL8 expression levels. Also, when compared with the WT strain, AE17ΔenvZ attenuated APEC pathogenicity in chickens. To explore the molecular mechanisms underpinning envZ in APEC17, we compared the WT and envZ-deletion strains using transcriptome analyses. RNA-Seq results identified 711 differentially expressed genes (DEGs) in the envZ mutant strain and DEGs were mainly enriched in outer membrane proteins, stress response systems, and TCSs. Quantitative real-time reverse transcription PCR (RT-qPCR) showed that EnvZ influenced the expression of biofilms and stress responses genes, including ompC, ompT, mlrA, basR, hdeA, hdeB, adiY, and uspB. We provided compelling evidence showing EnvZ contributed to APEC pathogenicity by regulating biofilms and stress response expression.

Acetoacetyl-CoA transferase ydiF regulates the biofilm formation of avian pathogenic Escherichia coli.

Avian pathogenic Escherichia coli (APEC) causes persistent infection of poultry and multi-system diseases, which seriously endanger the development of the poultry industry. Biofilm allows bacteria to adapt to the natural environment and plays an important role in resistance to the external environment and the pathogenicity of APEC, but the mechanism of its formation and regulatory network have not been clarified. In this study, we used a Tn5 transposon random mutation library constructed with APEC and identified ydiF, a gene that has not previously been recognized in E. coli biofilm formation. To confirm that the ydiF gene really can regulate the formation of APEC biofilm, the ydiF gene deletion strain was constructed using APEC81. Protein association networks prediction results show that ydiF is mainly associated with genes related to the metabolism of sugars and fatty acids. Deletion of the ydiF gene significantly reduces the formation of APEC biofilm and scanning electron microscopy indicated that the degree of adhesion between the bacteria was also reduced. The deletion of the ydiF gene also significantly reduced the motility of APEC81 and through transmission electron microscopy APEC81 was observed to have significantly fewer flagella. However, the colony morphology of APEC81 on Congo red and Coomassie brilliant blue media was unaffected. The results of fluorescence quantification showed that the deletion of the ydiF gene caused a down-regulation in the transcription of genes related to the second messenger, sugar metabolism, and quorum sensing. These results indicate that ydiF plays an important role in biofilm formation and the movement of APEC. In addition, it may be possible to regulate the formation of APEC biofilms by different methods such as by regulating the second messenger and metabolic system.

Cytokine Profiles and the Relationship of Disease Severity in Patients with Psoriasis.

Background: Psoriasis is a chronic skin disease characterized by hyperproliferation of keratinocytes and increased inflammation. Previous studies have detected the levels of cytokines in the serum of patients with psoriasis, yet few multi-cytokine combination studies have been reported. Objective: The aim of the study was to compare the levels of cytokines in the serum between patients with psoriasis and healthy controls, elucidate which factors influence the psoriasis progression. Methods: A total of 39 psoriasis patients and 30 healthy volunteers were enrolled. The venous blood was collected and the levels of 13 inflammatory cytokines were measured by human inflammation panel 1 kit. The severity of the disease was determined according to the psoriasis area and severity index (PASI) score. Results: Compared with healthy controls, the levels of nine cytokines (IFN-γ, TNF-α, IL-1ß, IL-6, IL-10, IL-12P40, IL-18, IL-17A and IL-23) were significantly increased, while the level of MCP-1 decreased in psoriatic patients. In addition, except for MCP-1, IL-10 and IL-12P40, these cytokine levels were positively correlated with the PASI score. Furthermore, there were higher serum lever of IFN-γ, TNF-α, IL-1ß, IL-6, IL-17A, IL-18 and IL-23 in active psoriasis than healthy controls and retrograde psoriasis. Conclusions: Increased serum levels of IFN-γ, TNF-α, IL-1ß, IL-6, IL-17A, IL-18 and IL-23 in psoriatic patients were associated with PASI and the stage of disease, which suggested that these cytokines play an important role in the pathogenesis of psoriasis. The detection of these cytokines can better observe the disease activity of psoriasis and optimize the treatment strategy.